Isolasi Protoplas secara Enzimatis pada Tanaman Kecipir

Imron Riyadi

Abstract


The
kind and concentration of enzyme that appropriate affected
isolation process and result of plant protoplast. The research
was conducted to enhance the protoplast rendements of winged
bean (Psophocarpus tetragonolobus L.) that was isolated by
Cellulase RS and Macerozyme R-10 enzyme as single and its
combination in a solution. Concentrations of enzyme used
were as much as 2.0-3.0% w/v for Cellulase RS and 0.4-0.6%
w/v for Macerozyme R-10. The solution containt mannitol 25
mM as an osmotycum. Isolation process was done on shaker
with 50 rpm (rotation per minute) speed in dark room for 5
hours. Results showed that C3 treatment (concentration of
Cellulase RS enzyme as much as 3.0% w/v) resulted
protoplasts density 3.49 x 105 protoplasts/g fresh weight of
hypocotyl and M2 treatment (concentration of Macerozyme R-
10 enzyme as much as 0.5% w/v) resulted 3.19 x 105
protoplasts/g. Whereas the best combined enzymes treatment
was achieved by C3M2 (combination between Cellulase RS as
much as 3.0% and Macerozyme R-10 enzyme as much as 0.5%
w/v) which resulted protoplasts density 4.94 x 105
protoplasts/g fresh weight of hypocotyl. The protoplast was
intact and viable.


Keywords


Psophocarpus tetragonolobus L.; protoplasts; isolation; rendement; density; Cellulase RS; Macerozyme R-10.

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DOI: http://dx.doi.org/10.21082/blpn.v16n1.2010.p57-63

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